Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study

Abstract

Earlier reports suggest that the endocannabinoids may play a role of endogenous tumor growth modulators. In this study, we investigated whether inhibition of the enzymes involved in the synthesis and degradation of endocannabinoids may reduce colorectal cancer cell invasion and migration. The human colon adenocarcinoma Colo-205 cells were incubated with PF-3845, JZL-184 and RHC-80267 (fatty acid amide hydrolase (FAAH), mono- (MAGL) and diacylglycerol lipase (DAGL) inhibitors, respectively) for 48 h. The MTT colorimetric assay was performed to quantify cell viability. Next, Colo-205 cells were incubated with PF-3845 alone or with PF-3845 together with selected antagonists: AM 251, AM 630, SB 366791, RN 1734 and G-15 (CB, CB, TRPV1, TRPV4 and GPR30 antagonists, respectively). Western blot assay was applied to identify the changes in CB and CB receptor expression. Migration and invasion assays were employed to characterize the effect of PF-3845 on colorectal cancer cell invasion. We found that of all the inhibitors used, the FAAH inhibitor PF-3845 reduced the Colo-205 cell line viability the most effectively (IC0=52.55 μM). We also showed that the effect of decreased cell viability was enhanced when Colo-205 cells were incubated with PF-3845 and RN-1734, a TRPV4 antagonist (IC0=30.54 μM). Western blot assay revealed significantly decreased CB receptor expression levels, while CB expression was increased in response to PF-3845 when compared to control. Furthermore, PF-3845 inhibited migration and invasion of Colo-205 cell line. These results suggest that pharmacological inhibition of FAAH and consequent enhancement of the endocannabinoid levels may reduce the colorectal cancer growth and progression.

Anticancer effects of n-3 EPA and DHA and their endocannabinoid derivatives on breast cancer cell growth and invasion

“…Our detection of FAAH in MCF-7 (hormone sensitive), but not in MDA-MB-231 (hormone insensitive) cells, agrees with our findings in prostate cells with similar hormone sensitivity status [10]. This seems to be different in colon cancer cells as a study by Wasilewski et al [22] showed that inhibiting FAAH with a selective inhibitor (PF-3845) decreased the viability, migration and invasiveness of the Colo-205 cell line. In contrast, the similar efficacies of endocannabinoids in inhibiting proliferation of both BC cell types observed in our study, irrespective of their high or low FAAH expression, suggested the enzyme was perhaps unimportant in this process or that sufficient endocannabinoid was always present in our in vitro system irrespective of FAAH activity.…”

Insights into the effects of the endocannabinoid system in cancer: a review

“…() postulated that the antiproliferative activity of AEA may be due to the reduction of polyamine levels that play a critical role in cell proliferation. Finally, the inhibition of FAAH also reduced the viability, migration and invasion of Colo‐205 cells in vitro (Wasilewski et al, ).…”

Physiological Role of Glutamate Dehydrogenase in Cancer Cells

“…Ammonium supplementation to glutamine-containing culture media was able to stimulate growth of the highly malignant HeLa, MDA-MB-231, PC3, HCT116, and Colo205 cancer cells in 2-D cultures. These human cancer cell lines are metastatic and hence have heightened abilities for migration, invasion, and colonization in glutamine-containing media (51)(52)(53)(54)(55)(56). They also show enhanced drug resistance due to overexpression of plasma membrane multidrug pumping ATPases, including P-glycoprotein and multidrug resistance protein-1 (57).…”

“…Cell growth was followed by counting cellular density every 24 h during 7 to 8 days. Viability was determined by the trypan blue assay, which revealed <10% cellular death (54). The duplication time was determined by using the following equation:…”